Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:2.7.7.49 (reverse transcriptase)
31,746 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The zinc-finger-Y (ZFY) gene is a candidate for the testis-determining-factor gene (TDF) on the human Y chromosome and is postulated to initiate testis differentiation during embryogenesis. However, the present study indicates that the ZFY gene and its X homologue (ZFX) are differentially expressed in adult tissues. A human testis-specific ZFY cDNA was isolated and completely sequenced. The corresponding ZFY transcript encodes a protein that has 801 amino acids and a calculated molecular weight of 90.6 kD. Expression analysis demonstrated that ZFY is transcribed primarily as 3- and 5.7-kb mRNA in testis and somatic cells, respectively. In contrast, the ZFX gene is expressed as a 5-kb transcript in the testis and as 6.7- and 8-kb transcripts in both ovarian and somatic tissues. With sets of gene-specific oligonucleotides, the origin and relative amount of the respective transcripts can be demonstrated in both Northern hybridization and reverse transcriptase-polymerase chain reaction analysis. Significantly, the 3-kb ZFY transcript was also detected in other mammalian adult testes. The testis-specific transcription of the ZFY gene hence suggests that it serves a conserved function in this organ.
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PMID:The putative testis-determining factor and related genes are expressed as discrete-sized transcripts in adult gonadal and somatic tissues. 251 51

Sry is the testes determining factor gene located on the Y chromosome. Expression of Sry had previously been found to occur during a short time frame of 10.5 to 12.5 days post coitum in the developing genital ridge. A recent study of the Sry transcript from adult testes found that the most abundant transcript is circular in nature. We have performed reverse transcription polymerase chain reactions with RNA from mouse preimplantation embryos ranging from the 2 cell stage to the blastocyst stage and report the detection of both circular and linear forms of Sry in the preimplantation embryo. In addition, we also demonstrate that the well described reverse transcriptase activity of Taq polymerase leads to a major difference in the mode of detection of the two forms.
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PMID:Detection of circular and linear transcripts of Sry in pre-implantation mouse embryos: differences in requirement for reverse transcriptase. 750 63

The micropia transposable element of Drosophila hydei is a long terminal repeat-containing retrotransposon present in both the autosomes and the Y chromosome. micropia expression gives rise to a complex set of sense and antisense RNAs transcribed primarily during spermatogenesis. The most abundant sense RNAs constitute an assortment of heterogeneous high-molecular-weight transcripts expressed as constituents of the Y-chromosomal lampbrush loops of primary spermatocytes. In addition, micropia encodes a full-length RNA that extends between the two long terminal repeats of the element. The major 1.0-kb antisense RNA characterized is complementary to the reverse transcriptase and RNase H coding regions of micropia. It is expressed from a testis-specific promoter during the primary spermatocyte stages and is detectable until spermatid elongation stages. Sequence comparison of this promoter with the 5' region of other testis-specific genes allows the conception of a conserved sequence that is responsible for this pattern of expression. A 284-bp fragment containing this sequence is able to drive testis-specific expression of the Escherichia coli lacZ gene in Drosophila melanogaster. This sequence is conserved in the micropia elements present in other Drosophila species that also encode an antisense RNA. The evolutionary conservation of micropia antisense RNA expression and the sequences responsible for its testis-specific transcription suggests a role for this antisense RNA in the control of germ line expression of the full-length transcript or transposon-encoded proteins.
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PMID:The Drosophila micropia retrotransposon encodes a testis-specific antisense RNA complementary to reverse transcriptase. 750 47

In trying to develop methods of gene therapy for Gaucher disease that will avoid the morbidity and mortality associated with bone marrow (BM) ablation, we transplanted BM stem cells transduced with a retroviral vector containing the human glucocerebrosidase cDNA into normal, nonablated, syngeneic mice. Donor BM from untreated male mice or treated with 5-fluorouracil (5-FU) was transduced ex vivo using a standard 4-day transduction protocol. Recipient female mice were injected one time only or once daily for 5 consecutive days or once a week for 5 consecutive weeks using 2 x 10(7) (untreated BM) or 2 x 10(6) (5-FU-treated BM) cells per injection. Initial transduction efficiency into colony-forming unit-spleen (CFU-S) was 80% to 100%. Recipient analysis was performed at least 6 months after the last transplantation. The best engraftment of donor stem cells, up to 5% by secondary CFU-S analysis, was obtained with multiple injections of transduced BM not previously treated with 5-FU. Polymerase chain reaction (PCR) amplification for both the transgene and the Y chromosome identified the progeny of transduced stem cells in various hematopoietic and non-hematopoietic organs. The copy number of the transgene in stem cells was 0.13 to 2.8. Transgene expression was shown by reverse transcriptase-PCR, in situ hybridization, and immunohistochemistry. No serious side effects of the procedure were noted. We conclude that multiple transplants of retrovirally transduced BM cells into nonablated recipients may be a safe and effective therapeutic modality for a number of genetic hematopoietic disorders.
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PMID:Transfer of the human glucocerebrosidase gene into hematopoietic stem cells of nonablated recipients: successful engraftment and long-term expression of the transgene. 762 Jan 75

The terminal portion of the short arm of the human Y (Yp) chromosome encodes a zinc-finger DNA binding protein (ZFY). A highly homologous gene, ZFX, is encoded on Xp. The potential of the zinc finger motif for regulating the expression of other genes suggests a role for this protein in the development of malignancy. Prostate adenocarcinoma is a malignancy of male-specific tissue, the incidence of which increases beyond the fifth decade of life. We have analyzed samples of human prostate adenocarcinoma for the expression of ZFY and ZFX transcripts. We found expression of ZFY transcripts in 3 of 31 prostate adenocarcinomas by using Northern analysis. No ZFY or ZFX transcripts were detected in normal hypertrophic prostate tissue on Northern analysis. In one prostate adenocarcinoma, high levels of the 5.1 kb ZFY and the 8.0 and 6.3 kb ZFX transcripts were present. In addition, this high-grade tumor contained a novel 4.3 kb transcript. When we used reverse transcriptase PCR (RT-PCR) to analyze these same samples, the number of tumors expressing ZFY and/or ZFX transcripts increased to 20 of 31. Transcripts for these genes were also present in the DU-145 and LNCaP human prostate adenocarcinoma cell lines. In 2 of the 6 benign prostatic hypertrophy (BPH) tissues analyzed by RT-PCR, barely detectable products of ZFY were observed, and none contained ZFX products. Southern analysis revealed that the portion of the Y chromosome which contains the ZFY gene was not lost from the majority of the tumor cells in any of the prostate malignancies examined.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:ZFY gene expression and retention in human prostate adenocarcinoma. 768 Aug 90

Because male ovine embryos develop faster than female embryos, the transcription of SRY and ZFY, two genes located on the Y chromosome, was examined in preimplantation stages using the reverse transcriptase polymerase chain reaction (RT-PCR). RNA was extracted from pools of ovine embryos matured and fertilized in vitro then cultured in synthetic oviduct fluid medium and recovered from 24 to 207 hr post-insemination (two-cell up to hatched blastocyst stage). Since primers used to amplify ZFY also amplify the homologue ZFX, located on the X chromosome, transcripts were differentiated by digestion with restriction enzymes. ZFY and ZFX transcripts were present in all stages examined following RT-PCR, whereas transcripts for SRY were undetectable in all investigated stages following either RT nested PCR or Southern analysis. The presence of ZFY transcripts suggests that Y chromosome is transcriptionally active during early ovine preimplantation development. The possible relationship between a faster growth of male embryos and the transcription of Y-linked genes at early stages of development is discussed.
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PMID:Transcription of Y- and X-linked genes in preimplantation ovine embryos. 891 69

We have examined mRNA expression of two genes located on the Y chromosome, the sex-determining region Y gene (SRY) and the linked zinc finger gene (ZFY), using in vitro fertilized-in vitro cultured bovine embryos. Expression of the SRY gene, implicated in sex determination in mammals, has been reported to occur both for a short time at the sex-determining stage of development around the period of the primitive undifferentiated gonad and in the adult testis. In this study, using a sensitive reverse transcriptase-polymerase chain reaction (RT-PCR) assay, we detected SRY but not ZFY mRNA expression as early as the 4- to 8-cell stage and through to the blastocyst stage in bovine embryos. The expression of SRY at these early stages and the previous observation that in vitro-produced male bovine embryos develop faster in culture than female embryos suggest that sex differences are evident prior to gonadal differentiation and that preimplantation bovine embryos have sexually dimorphic gene expression at least with respect to SRY transcripts.
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PMID:Early transcription of the SRY gene by bovine preimplantation embryos. 929 74

Random amplification of polymorphic DNA (RAPD) was used to analyse genomic DNA from virgin females and males of Brugia malayi, with a view to identifying sex-specific differences predicted by an XX/XY system of chromosomal sex determination. A product of 2338 bp, amplified with the arbitrary primer 5' GTTGCGATCC 3', was obtained exclusively from males. Primers based on the sequence of this product amplified a DNA fragment of the expected size from each of two independent isolates of B. malayi (from Malaysia and Indonesia) by PCR. No reaction product was obtained from the closely related species Brugia pahangi. In a genetic cross between B. malayi males and B. pahangi females, F1 hybrid microfilariae were PCR-positive, indicating that the locus is paternally-inherited. Southern blotting demonstrated that the target sequence resides in the high molecular weight fraction of genomic DNA, confirming that it is of chromosomal, rather than mitochondrial, origin. Sequencing of the locus revealed significant similarity with members of a family of reverse transcriptase-like genes in Caenorhabditis elegans. In-frame stops indicate that the gene is non-functional, but multiple bands of hybridisation in Southern blots suggest that the RT sequence may be the relic of a transposable element. Multiple repeats of the dinucleotide AT occurred in another region of the sequence. These varied in number between the two isolates of B. malayi in the manner of a microsatellite, surprisingly the first to be described from the B. malayi genome. Because of its association with the Y chromosome, we have given the locus the acronym TOY (Tag On Y). Identification of this chromosome-specific marker confirms the XX/XY heterogametic karyotype in B. malayi and opens the way to elucidation of the role of Y in sex determination.
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PMID:Identification of a molecular marker for the Y chromosome of Brugia malayi. 1021 19

Male-associated DNA sequences were analysed in hemp (Cannabis sativa L.), a dioecious plant with heteromorphic sex chromosomes. A male-associated DNA sequence in C. sativa (MADC1) and its flanking sequence encoded a reverse transcriptase that was strongly homologous to those of LINE-like retrotransposons from various plants and other organisms, as well as another open reading frame (ORF). Fluorescence in situ hybridization (FISH) with MADC1 as probe, which yielded strong signals specific for male genomic DNA in gel blot analysis, generated a clear doublet signal at the end of the long arm of the Y chromosome. FISH using pachytene chromosomes of pollen mother cells at meiotic prophase I revealed that pairing of X and Y chromosomes occurred at the short arm of the Y chromosome where MADC1 was not present. Furthermore, FISH using extended DNA fibers, with MADC1 and its flanking DNA as probes, revealed that 100 to 200 copies of the retrotransposon were located in tandem on the Y chromosome. These results support the hypothesis that accumulation of a specific LINE-like retrotransposon at the terminal region of the long arm of the Y chromosome might be one cause of heteromorphism of sex chromosomes.
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PMID:Site-specific accumulation of a LINE-like retrotransposon in a sex chromosome of the dioecious plant Cannabis sativa. 1120 35

We have isolated a new interspersed sequence present in a high copy number in the ovine genome. This patchwork sequence, named 3.79 AS1, is part of a larger element encompassing similarities to constant region of reverse transcriptase and to art2 shared with the Bovine Dimer Driven Family (BDDF). The 3.79 AS1 sequence includes homologies to amplification promoting sequences (APS), to a potential origin of bidirectional DNA replication (OBR), to the Alu core sequence motif GGAGGC required for RNA polymerase III promoter function and to the ATGGCTGCCAT sequence that has been shown to be able to induce amplification-dependent transformation in murine cells. Fluorescent in situ hybridization experiments using probes derived from both ends of the 3.79 AS1 sequence showed a widespread signal over all sheep chromosomes, except the Y chromosome. We propose that the structural features of the 3.79 AS1 patchwork sequence, that is likely to be a subfamily of Bov B LINE that invaded the Artiodactyl genome prior to the separation of the Bovidae species, facilitated its massive amplification and dispersion in the ovine genome.
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PMID:A patchwork interspersed sequence is present in a high copy number in the sheep genome. 1255 68


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